Electrophysiological Analysis of Synaptic Transmission in Central Neurons of Drosophila Larvae

Department of Biology, University of Utah, Salt Lake City, Utah 84112 Rohrbough, Jeffrey and Kendal Broadie. Electrophysiological Analysis of Synaptic Transmission in Central Neurons of Drosophila Larvae. J. Neurophysiol. 88: 847-860, 2002. We report functional neuronal and synaptic transmission pro...

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Published in:Journal of neurophysiology 2002-08, Vol.88 (2), p.847-860
Main Authors: Rohrbough, Jeffrey, Broadie, Kendal
Format: Article
Language:English
Subjects:
3',5'-Cyclic-AMP Phosphodiesterases - genetics
Acetylcholine - metabolism
Acetylcholine - pharmacology
Action Potentials - drug effects
Animals
Calcium - metabolism
Central Nervous System - physiology
Drosophila
Drosophila Proteins
Dynamins
Electric Stimulation
Electrophysiology
GABA Antagonists - pharmacology
gamma-Aminobutyric Acid - metabolism
gamma-Aminobutyric Acid - pharmacology
Glutamic Acid - metabolism
Glutamic Acid - pharmacology
GTP Phosphohydrolases - genetics
Mutation
Neurons - physiology
Nicotinic Antagonists - pharmacology
Patch-Clamp Techniques
Picrotoxin - pharmacology
Receptors, Nicotinic - metabolism
Synaptic Transmission - drug effects
Synaptic Transmission - physiology
Tubocurarine - pharmacology
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Summary:Department of Biology, University of Utah, Salt Lake City, Utah 84112 Rohrbough, Jeffrey and Kendal Broadie. Electrophysiological Analysis of Synaptic Transmission in Central Neurons of Drosophila Larvae. J. Neurophysiol. 88: 847-860, 2002. We report functional neuronal and synaptic transmission properties in Drosophila CNS neurons. Whole cell current- and voltage-clamp recordings were made from dorsally positioned neurons in the larval ventral nerve cord. Comparison of neuronal Green Fluorescent Protein markers and intracellular dye labeling revealed that recorded cells consisted primarily of identified motor neurons. Neurons had resting potentials of 50 to 60 mV and fired repetitive action potentials (APs) in response to depolarizing current injection. Acetylcholine application elicited large excitatory responses and AP bursts that were reversibly blocked by the nicotinic receptor antagonist D -tubocurarine (dtC). GABA and glutamate application elicited similar inhibitory responses that reversed near normal resting potential and were reversibly blocked by the chloride channel blocker picrotoxin. Multiple types of endogenous synaptically driven activity were present in most neurons, including fast spontaneous synaptic events resembling unitary excitatory postsynaptic currents (EPSCs) and sustained excitatory currents and potentials. Sustained forms of endogenous activity ranged in amplitude from smaller subthreshold "intermediate" sustained events to large "rhythmic" events that supported bursts of APs. Electrical stimulation of peripheral nerves or focal stimulation of the neuropil evoked sustained responses and fast EPSCs similar to endogenous events. Endogenous activity and evoked responses required external Ca 2+ and were reversibly blocked by dtC application, indicating that cholinergic synaptic transmission directly underlies observed activity. Synaptic current amplitude and frequency were reduced in shibire conditional dynamin mutants and increased in dunce cAMP phosphodiesterase mutants. These results complement and advance those of recent functional studies in Drosophila embryonic neurons and demonstrate the feasibility of in-depth synaptic transmission and plasticity studies in the Drosophila CNS.
ISSN:0022-3077
1522-1598
DOI:10.1152/jn.2002.88.2.847